Preparation method and application of spirulina nanoemulsion

Abstract

The invention discloses a preparation method and application of spirulina nanoemulsion, and belongs to the technical field of functional foods.The preparation method comprises the following steps that S1, a water phase is prepared, specifically, fresh spirulina mud is taken, washed and filtered, and a spirulina mud raw material is obtained; adding purified water of which the weight is 2-3 times that of the algae mud raw material into the algae mud raw material, and then carrying out superfine shearing treatment; centrifuging after the shearing treatment is finished, filtering and collecting filtrate to obtain a water phase; s2, preparing an oil phase: weighing egg yolk lecithin and coconut oil according to a mass ratio of 1: (0.5-0.8), and uniformly mixing and stirring the egg yolk lecithin and the coconut oil to obtain the oil phase; s3, preparing a compound emulsifier, wherein the compound emulsifier comprises the following components: tween-80, span-80, chicoric acid and 1, 2-propylene glycol in a mass ratio of (3-4): (5-7): (1-1.5): (4-5); and S4, emulsifying: mixing the water phase and the oil phase according to the mass ratio of 1: (0.5-1), adding a compound emulsifier which is 2-3 times of the mass of the mixture, and uniformly stirring to obtain the spirulina nanoemulsion. The spirulina nanoemulsion prepared by the preparation method disclosed by the invention has the effect of high stability.

Description

Technical Field

[0001] The present invention belongs to the technical field of functional foods. Specifically, it relates to a preparation method and application of spirulina nanoemulsion. Background Art

[0002] Spirulina is an ideal human food discovered by current scientific research and is praised by some scholars as the "nutrition champion" and "future food". Its extracts have been recognized in the field of anti-cancer drug research and development. In addition, spirulina also has the effect of assisting in treating diseases and is highly regarded by scientific researchers. It is the most nutritionally balanced and comprehensive natural food discovered so far. Spirulina contains a variety of bioactive substances, such as phycocyanin, phenols, polysaccharides, polyunsaturated fatty acids (PUFAs), carotenoids, vitamins, and sterols. Most of these compounds play an important role in the treatment of cardiovascular diseases (CVDs), high cholesterol, high blood sugar, obesity, hypertension, tumors, and inflammatory diseases and are considered a natural medicine.

[0003] Commercially available spirulina products are mostly made into tablets and capsules with spirulina dry powder as the raw material. They have a large dosage and are not convenient for some people with difficulty swallowing. Most of the components with the effects of enhancing immunity, promoting metabolism, and antioxidation in spirulina are water-soluble components. Separating them and preparing them into nanoemulsion can improve the stability and bioavailability of the active ingredients and is more conducive to the use of people with special needs.

[0004] Some existing nanoformulations are prepared by extracting a single component from spirulina dry powder, and the utilization rate of the active ingredients in spirulina is not high. The present invention uses fresh spirulina as the raw material, obtains the water-soluble components therein, and prepares them into nanoemulsion after concentration, which can retain the nutritional components of spirulina to the greatest extent and has the effects of high stability and helping to enhance immunity. Summary of the Invention

[0005] In order to overcome the problems existing in the prior art, the present invention provides a preparation method and application of spirulina nanoemulsion.

[0006] To achieve the above object, the present invention is realized by the following technical solutions: The first aspect of the present invention provides a preparation method of spirulina nanoemulsion, which comprises the following steps: S1, preparation of aqueous phase: Take fresh spirulina mud, wash and filter it to obtain the algal mud raw material; add purified water with a weight 2-3 times that of the algal mud raw material thereto and then perform ultra-fine shearing treatment; after the shearing treatment is completed, centrifuge and filter to collect the filtrate to obtain the aqueous phase; S2, preparation of oil phase: Weigh egg yolk lecithin and coconut oil according to a mass ratio of 1:0.5-0.8, and then mix and stir them evenly to obtain the oil phase; S3. Preparation of compound emulsifier: The compound emulsifier comprises components in the following mass ratio: Tween-80: Span-80: chicoric acid: 1,2-propanediol = 3-4: 5-7: 1-1.5: 4-5; S4. Emulsification: After mixing the aqueous phase and the oil phase in a mass ratio of 1: 0.5-1, add a compound emulsifier 2-3 times the mass of the mixture, stir evenly, and sterilize with an electric pulse sterilization device to obtain the spirulina nanoemulsion.

[0007] Further, in the step S1, the spirulina mud is washed and filtered to contain 40%-50% water.

[0008] Further, in the step S1, the method of ultrafine shearing treatment is: use a food ultrafine shearing machine to treat for 8-9 h, and adopt an intermittent working method during the treatment process, shear for 30 s and stop for 30 s.

[0009] Further, the compound emulsifier comprises components in the following mass ratio: Tween-80: Span-80: chicoric acid: 1,2-propanediol = 3.5: 6: 1.3: 4.5.

[0010] The second aspect of the present invention provides a spirulina nanoemulsion, and its preparation process is the preparation method described in the first aspect.

[0011] The third aspect of the present invention provides the application of the nanoemulsion in the first aspect in functional foods, drugs or drug additives.

[0012] Through the above technical solutions, the present invention can at least achieve the following beneficial effects: The spirulina nanoemulsion prepared by the preparation method of the present invention has the effects of high stability and helping to enhance immunity. Specific embodiments

[0013] Unless otherwise specified, the materials and reagents used in the present invention are commercially available. Example 1

[0014] A preparation method of a spirulina nanoemulsion comprises the following steps: S1. Preparation of aqueous phase: Take fresh spirulina mud, wash and filter it to contain 40%-50% water to obtain algal mud raw material; add purified water 2.5 times its weight to the algal mud raw material and then carry out ultrafine shearing treatment for 8.5 h; after the shearing treatment, centrifuge and filter to collect the filtrate to obtain the aqueous phase; wherein, in order to prevent protein denaturation, an intermittent working method is adopted during the ultrafine shearing treatment, shear for 30 s and stop for 30 s.

[0015] S2. Preparation of oil phase: Weigh egg yolk lecithin and coconut oil in a mass ratio of 1: 0.6, and then mix and stir them evenly to obtain the oil phase; S3. Preparation of compound emulsifier: The compound emulsifier contains components in the following mass ratio: Tween-80: Span-80: chicoric acid: 1,2-propanediol = 3.5: 6: 1.3: 4.5.

[0016] S4. Emulsification: Mix the aqueous phase and the oil phase in a mass ratio of 1:0.7, then add a compound emulsifier that is 2.5 times the mass of the mixture, stir evenly, and after sterilization treatment with an electric pulse sterilization device, the spirulina nanoemulsion is obtained. Example 2

[0017] A method for preparing a spirulina nanoemulsion comprises the following steps: S1. Preparation of aqueous phase: Take fresh spirulina mud, wash and filter it until the water content is 40%-50% to obtain the algal mud raw material; add purified water that is 2 times its weight to the algal mud raw material and then carry out ultra-fine shearing treatment for 8 h; after the shearing treatment is completed, centrifuge and filter to collect the filtrate to obtain the aqueous phase; among them, in order to prevent protein denaturation, an intermittent working method is adopted during the ultra-fine shearing treatment, shearing for 30 s and stopping for 30 s.

[0018] S2. Preparation of oil phase: Weigh egg yolk lecithin and coconut oil according to a mass ratio of 1:0.5, and then mix and stir them evenly to obtain the oil phase; S3. Preparation of compound emulsifier: The compound emulsifier contains components in the following mass ratio: Tween-80: Span-80: chicoric acid: 1,2-propanediol = 3: 5: 1: 4.

[0019] S4. Emulsification: Mix the aqueous phase and the oil phase in a mass ratio of 1:0.5, then add a compound emulsifier that is 2 times the mass of the mixture, stir evenly, and after sterilization treatment with an electric pulse sterilization device, the spirulina nanoemulsion is obtained. Example 3

[0020] A method for preparing a spirulina nanoemulsion comprises the following steps: S1. Preparation of aqueous phase: Take fresh spirulina mud, wash and filter it until the water content is 40%-50% to obtain the algal mud raw material; add purified water that is 3 times its weight to the algal mud raw material and then carry out ultra-fine shearing treatment for 9 h; after the shearing treatment is completed, centrifuge and filter to collect the filtrate to obtain the aqueous phase; among them, in order to prevent protein denaturation, an intermittent working method is adopted during the ultra-fine shearing treatment, shearing for 30 s and stopping for 30 s.

[0021] S2. Preparation of oil phase: Weigh egg yolk lecithin and coconut oil according to a mass ratio of 1:0.8, and then mix and stir them evenly to obtain the oil phase; S3. Preparation of compound emulsifier: The compound emulsifier contains components in the following mass ratio: Tween-80: Span-80: chicoric acid: 1,2-propanediol = 4: 7: 1.5: 5 S4, Emulsification: Mix the aqueous phase and the oil phase in a mass ratio of 1:1, then add a compound emulsifier three times the mass of the mixture, stir evenly, and after sterilization treatment with an electric pulse sterilization device, the spirulina nanoemulsion is obtained. Comparative Example 1

[0022] The difference between Comparative Example 1 and Example 1 is that the compound emulsifier does not contain chicoric acid. Stability Test

[0023] 1. High-speed Centrifugation Test

[0024] Take the nanoemulsions prepared in Examples 1 to 3 and Comparative Example 1, centrifuge at a speed of 12,000 r / min for 30 min, observe the properties of the nanoemulsions, and the statistics are shown in Table 1 below.

[0025] Table 1 High-speed Centrifugation Test Table of Nanoemulsions Prepared by Different Preparation Methods

[0026] 2. Accelerated Test Test Scheme Design: According to the guiding principles for the stability test of bulk drugs and drug preparations (Part 4 of the Chinese Pharmacopoeia 2020 Edition, 9001), take 100 g of the nanoemulsions prepared in Examples 1 to 3 and Comparative Example 1 respectively. Place them in clean glass bottles, set the temperature at 40°C ± 2°C and the relative humidity at 75% ± 5%, and place them for 6 months. Sampling is carried out at 0 month, 1 month, 2 months, 3 months, and 6 months respectively. The test results are shown in Table 2 below: Table 2 Statistical Table of Accelerated Test Results 3. Immune Enhancement Experiment

[0027] 3.1 Effects of the nanoemulsions prepared in each example and comparative example on the activity of NK cells in mice

[0028] Take the nanoemulsions prepared in Examples 1 to 3 and Comparative Example 1. Take 130 healthy mice, 10 in each group, divide them into 13 groups, with three groups for each treatment in Examples 1 to 3 and Comparative Example 1, divided into high, medium, and low dose groups, and set one blank control group. Administer the drugs by gavage according to the dosage in Table 3 below, and adjust the dosage according to the change in the body weight of the animals every week. The mice are sacrificed 30 days after drug administration, and the spleens are removed under sterile conditions. Grind the removed spleens and pass them through a 200-mesh sieve to make a single cell suspension, and adjust the cell concentration with complete culture medium. Add 100 μL of target cells and spleen lymphocytes to a 96-well plate respectively. After completing the operation according to the procedure, use an enzyme-labeled instrument to measure the optical density value (OD) at 490 nm and calculate the activity of NK cells. The results are shown in Table 4.

[0029] Table 3 Dosage Table of Nanoemulsions Prepared in Each Example and Comparative Example

[0030] Table 4 Effects of nanoemulsions prepared in each example and comparative example on the activity of NK cells in mice

[0031] As can be seen from Table 4 above, the activity of NK cells in the test sample groups (Examples 1 to 3) was significantly higher than that in the control group and Comparative Example 1. 3.2 Effects of nanoemulsions prepared in each example and comparative example on the carbon clearance index of mice

[0032] Take the nanoemulsions prepared in Examples 1 to 3 and Comparative Example 1. Take 130 healthy mice, 10 in each group, divided into 13 groups. Each of Examples 1 to 3 and Comparative Example 1 has three groups for each treatment, divided into high, medium, and low dose groups, and one group is set as the blank control. Administer the drugs by gavage according to the dose in Table 3 below, and adjust the dosage according to the change in the animal's body weight per week. The duration of the entire experiment is 14 days. 30 minutes after the last drug administration, inject 0.1 ml / 10 g body weight of Indian ink into the tail vein of each mouse. Take 20 μl of blood from the orbital vein of the mouse at 1 minute and 5 minutes respectively, add it to 2 ml of 0.1% NaCO3 solution and shake well, then measure the optical density (OD) by colorimetry at 680 nm in a spectrophotometer, and calculate the clearance index (K). The results are shown in Table 5.

[0033] Table 5 Effects of nanoemulsions prepared in each treatment on the carbon clearance index of mice

[0034] As can be seen from Table 5 above, the carbon clearance index of the test sample groups (Examples 1 to 3) was significantly higher than that of the control group and Comparative Example 1.

[0035] In summary, the nanoemulsions prepared in Examples 1 to 3 of this application help to enhance immunity.

[0036] Finally, it should be noted that the above preferred embodiments are only used to illustrate the technical solutions of the present invention and are not restrictive. Although the present invention has been described in detail through the above preferred embodiments, those skilled in the art should understand that various changes can be made in form and details without departing from the scope defined by the claims of the present invention.

Claims

1. A preparation method of spirulina nanoemulsion, characterized in that: It comprises the following steps: S1, preparation of aqueous phase: Take fresh spirulina mud, wash and filter it to obtain the algal mud raw material; add purified water which is 2 - 3 times the weight of the algal mud raw material thereto, and then perform ultra-fine shearing treatment; after the shearing treatment, centrifuge and filter to collect the filtrate to obtain the aqueous phase. S2, preparation of oil phase: Weigh lecithin and coconut oil according to the mass ratio of 1:0.5 - 0.8, and then mix and stir them evenly to obtain the oil phase. S3, preparation of compound emulsifier: The compound emulsifier comprises components with the following mass ratio: Tween - 80: Span - 80: chicoric acid: 1,2 - propanediol = 3 - 4: 5 - 7: 1 - 1.5: 4 - 5; S4, emulsification: Mix the aqueous phase and the oil phase according to the mass ratio of 1:0.5 - 1, then add a compound emulsifier which is 2 - 3 times the mass of the mixture, stir evenly, and after sterilization treatment with an electric pulse sterilization device, the spirulina nanoemulsion is obtained.

2. The preparation method of a spirulina nanoemulsion according to claim 1, wherein: In the step S1, the spirulina mud is washed and filtered until the water content is 40% - 50%.

3. The preparation method of a spirulina nanoemulsion according to claim 1, characterized in that: In the step S1, the method of ultra-fine shearing treatment is: use a food ultra-fine shearing machine to treat for 8 - 9 h, and during the treatment process, adopt an intermittent working method, shear for 30 s and stop for 30 s.

4. The preparation method of a spirulina nanoemulsion according to claim 1, characterized in that: The compound emulsifier comprises components with the following mass ratio: Tween - 80: Span - 80: chicoric acid: 1,2 - propanediol = 3.5: 6: 1.3: 4.

5.

5. A Spirulina nanoemulsion, characterized in that: Its preparation method is any one of claims 1 - 4.

6. Use of the nanoemulsion as described in claim 5 in functional foods, drugs or pharmaceutical additives.

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